Team:Calgary/Sandbox/Notebook/Protocols/AgaroseGelElectrophoresis

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<h1>Agarose Gel Electrophoresis</h1>
<h1>Agarose Gel Electrophoresis</h1>
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<p><b>Reagents and Materials</b></p>
+
<h2>Reagents and Materials<h2>
-
  <p>1X TA </p>
+
<ul>
-
    <p>Graduated Cylinder</p>
+
<li>1X TAE Buffer</li>
-
    <p>125 mL flask </p>
+
<li>Graduated Cylinder</li>
-
    <p>Agarose </p>
+
<li>125 mL flask</li>
-
    <p>Gel Pouring Tray </p>
+
<li>Agarose</li>
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    <p>Tape </p>
+
<li>Gel Pouring Tray</li>
-
    <p>Gel rig </p>
+
<li>Tape</li>
-
    <p>Red Safe </p>
+
<li>Gel rig</li>
-
  <p><b>Protocol</b></p>
+
<li>Red Safe</li>
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    <p>Measure out 100mL of 1x TAE buffer;</p>
+
</ul>
-
    <p>Transfer buffer to 125 mL flask;</p>
+
<h2>Protocol</h2>
-
    <p>Weigh out enough agarose to make a 1% gel (in our case 1.0 g of agarose was the right amount);</p>
+
<ol>
-
    <p> Transfer agarose to 125mL flask;</p>
+
<li>Measure out 100mL of 1x TAE buffer</li>
-
    <p> Melt agarose in microwave until solution is almost boiling, stirring every 15-20 seconds (should be around 2 minutes);</p>
+
<li>Transfer buffer to 125 mL flask</li>
-
    <p> Allow agarose to cool (do not let it cool to the point where it is hard);</p>
+
<li>Weigh out enough agarose to make a 1% gel (in our case 1.0 g of agarose was the right amount)</li>
-
    <p> Add 5 uL of Red Safe to the cooling agarose;</p>
+
<li>Transfer agarose to 125mL flask</li>
-
    <p> Assemble the gel pouring apparatus by inserting gate into slots;</p>
+
<li>Melt agarose in microwave until solution is almost boiling, stirring every 15-20 seconds (should be around 2 minutes)</li>
-
    <p> Allow gel to cool until flask can be handled comfortably;</p>
+
<li>Allow agarose to cool (do not let it cool to the point where it is hard)</li>
-
    <p> Place comb in the gel rig;</p>
+
<li>Add 5 uL of Red Safe to the cooling agarose</li>
-
    <p> Pour agarose into gel tray;</p>
+
<li>Assemble the gel pouring apparatus by inserting gate into slots</li>
-
    <p> Allow to solidify. While the gel is solidifying prepare the samples. Add your sample and 1 uL 10x Loading Dye, 4 uL of DNA and 5 uL of water;</p>
+
<li>Allow gel to cool until flask can be handled comfortably</li>
-
    <p> Pour 1X TAE over gel so that gel is covered by a 3-5mm buffer;</p>
+
<li>Place comb in the gel rig</li>
-
    <p> Load samples into lane (Don't forget to load a 1kb+ ladder into one of the lanes);</p>
+
<li>Pour agarose into gel tray</li>
-
    <p> Hook electrodes to gel apparatus;</p>
+
<li>Allow to solidify. While the gel is solidifying prepare the samples. Add your sample and 1 uL 10x Loading Dye, 4 uL of DNA and 5 uL of water</li>
-
    <p> Run the apparatus at 100V for 30 - 45 minutes (make sure to watch that the dye does not run off the gel);</p>
+
<li>Pour 1X TAE over gel so that gel is covered by a 3-5mm buffer</li>
-
    <p> Visualize the gel and record the results.</p>
+
<li>Load samples into lane (Don't forget to load a 1kb+ ladder into one of the lanes)</li>
 +
<li>Hook electrodes to gel apparatus</li>
 +
<li>Run the apparatus at 100V for 30 - 45 minutes (make sure to watch that the dye does not run off the gel)</li>
 +
<li>Visualize the gel and record the results</li>
 +
</ol>
</section>
</section>
</html>
</html>

Latest revision as of 23:27, 17 September 2013

Agarose Gel Electrophoresis

Reagents and Materials

  • 1X TAE Buffer
  • Graduated Cylinder
  • 125 mL flask
  • Agarose
  • Gel Pouring Tray
  • Tape
  • Gel rig
  • Red Safe

Protocol

  1. Measure out 100mL of 1x TAE buffer
  2. Transfer buffer to 125 mL flask
  3. Weigh out enough agarose to make a 1% gel (in our case 1.0 g of agarose was the right amount)
  4. Transfer agarose to 125mL flask
  5. Melt agarose in microwave until solution is almost boiling, stirring every 15-20 seconds (should be around 2 minutes)
  6. Allow agarose to cool (do not let it cool to the point where it is hard)
  7. Add 5 uL of Red Safe to the cooling agarose
  8. Assemble the gel pouring apparatus by inserting gate into slots
  9. Allow gel to cool until flask can be handled comfortably
  10. Place comb in the gel rig
  11. Pour agarose into gel tray
  12. Allow to solidify. While the gel is solidifying prepare the samples. Add your sample and 1 uL 10x Loading Dye, 4 uL of DNA and 5 uL of water
  13. Pour 1X TAE over gel so that gel is covered by a 3-5mm buffer
  14. Load samples into lane (Don't forget to load a 1kb+ ladder into one of the lanes)
  15. Hook electrodes to gel apparatus
  16. Run the apparatus at 100V for 30 - 45 minutes (make sure to watch that the dye does not run off the gel)
  17. Visualize the gel and record the results

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